桃蛀螟对青霉菌侵染苹果的产卵选择与行为趋向反应

为探明桃蛀螟Conogethes punctiferalis偏好于在青霉菌Penicillium侵染的苹果上产卵的特点与机理,从苹果、玉米和柑橘上分离、纯化获得15株青霉菌,通过形态学和rDNA-ITS序列分析鉴定后,以健康苹果为对照,测试不同青霉菌菌株侵染的苹果对桃蛀螟产卵选择的影响,并利用四臂嗅觉仪测试桃蛀螟对青霉菌诱导的苹果挥发物的行为趋向反应。结果表明,15株青霉菌分别属于皮落青霉P. crustosum、橘青霉P. citrinum、苏门答腊青霉P. sumatrense和指状青霉P. digitatum的不同株型;桃蛀螟对其中的12株青霉菌侵染的苹果的产卵选择率均极显著高于健康苹果,尤其对皮落青霉CO5菌株、苏门答腊青霉OR3菌株和指状青霉OR5菌株侵染的苹果的产卵选择率最高,分别达到67.73%、61.50%和68.39%;桃蛀螟对感染皮落青霉CO5菌株和指状青霉OR5菌株的苹果挥发物的选择率分别为31.37%和29.19%,均显著高于对感染橘青霉OR1菌株和苏门答腊青霉OR3菌株的苹果挥发物的选择率18.14%和21.31%,且前二者之间、后二者之间均无显著差异。表明青霉菌可通过影响寄主植物挥发物释放,从而影响桃蛀螟对寄主植物的产卵选择和行为趋向反应。     

The Interaction of Endochitinase, A Synthetic Peptide and Resveratrol in Controlling Fungi in vitro

The efficacy of two-way combinations of Trichoderma endochitinase (CHIT42), synthetic peptide ESF12 and resveratrol upon the control of growth of Botrytis cinerea and Penicillium digitatum were evaluated in vitro. CHIT42 had effective concentration for 50% growth inhibition (EC₅₀) values of 1.5 and 0.5 µM against B. cinerea and P. digitatum, respectively. CHIT42 also caused swelling, hyperbranching and bursting of P. digitatum at concentrations as low as 0.003 µM. The EC₅₀ values of ESF12 were 30.9 and 29.2 µM against B. cinerea and P. digitatum, respectively. Resveratrol had EC₅₀ values of 178.6 and 343.2 µM against B. cinerea and P. digitatum, respectively. Combinations of CHIT42 + ESF12, CHIT42 + resveratrol and ESF12 + resveratrol showed additive activity (interaction ratios between 0.5 and 1.5) against B. cinerea and P. digitatum.     

Induction of Resistance to Penicillium digitatum in Grapefruit by β-Aminobutyric Acid

-Aminobutyric acid (BABA), an inducer of pathogen resistance in plants, induced disease resistance in reproductive parts of the plant, such as grapefruit peel tissue. Application of BABA to specific wound sites on the fruit peel surface induced resistance to Penicillium digitatum, the main postharvest pathogen of citrus fruit, in a concentration-dependent manner, being most effective at 20mM, and rather less effective at either higher or lower concentrations. The effect of BABA in inducing resistance to P. digitatum in the fruit peel surface was local and limited to the vicinity (within 1–2cm) of the BABA-treated site. In addition to inducing pathogen resistance, increasing concentrations of BABA (from 1 to 100mM) also exhibited direct antifungal activity and inhibited P. digitatum spore germination and germ tube elongation in vitro. The induction of resistance to P. digitatum by BABA was accompanied by the activation of various pathogen defense responses in grapefruit peel tissue, including activation of chitinase gene expression and protein accumulation after 48h, and an increase in phenylalanine ammonia lyase (PAL) activity after 72h.     

Studies on the selective fungitoxicity of aliphatic amines

A comparison of the uptake of simple (C₁–C₄) aliphatic amines byPenicillium digitatum revealed that the fungitoxicity of (–)sec-butylamine [(–)SBA] was not due to its accumulation by hyphae since (+)SBA was accumulated to the same extent and methylamine, which showed negligible antifungal activity, accumulated to twice the level of SBA. Amines with a secondary alkyl structure were resistant to fungal metabolism, whereas primary amines were degraded to a significant extent byP. digitatum during a 4 h incubation period.(–)SBA accumulated in the fresh hyphae ofP. digitatum to a level 24 times higher than that in the culture medium containing 1 mol ml^–1; most of the SBA effluxed from the hyphae when transferred to fresh culture medium minus SBA. SBA did not accumulate when hyphae were incubated in N₂ or in the presence of respiratory inhibitors. The absorption and accumulation of SBA is characteristic of active transport.     

Improving Control of Green and Blue Molds of Oranges by Combining Pantoea Agglomerans (CPA-2) and Sodium Bicarbonate

The potential of using Pantoea agglomerans (strain CPA-2) alone, or in combination with sodium bicarbonate or sodium carbonate solutions, for control of Penicillium digitatum (green mold) and Penicillium italicum (blue mold) on oranges was investigated under ambient (20 °C) and cold storage (3 °C) conditions. P. agglomerans controlled both pathogens on oranges at 2 × 10⁸ cfu ml^-1. The biocontrol agent was found to be completely tolerant to 2% sodium bicarbonate at room temperature, although its culturability was reduced by > 1000-fold after 30 min in 2% sodium carbonate. The efficacy of P. agglomerans for control of green mold was improved when combined with sodium bicarbonate, resulting in complete and 97.6% reduction of decay incidence at 3 °C and 20 °C, when compared to untreated controls. Satisfactory results were also obtained with the combined treatment for control of blue mold. P. agglomerans grew well inside wounds on oranges at both 20 °C and 3 °C. In contrast, it showed a reduced growth on the surface of intact fruit. Sodium bicarbonate at 2% concentration did not noticeably affect antagonist population development. Thus, use of bicarbonate treatment at 2% followed by the antagonist P. agglomerans CPA-2 could be an alternative to chemicals for control of postharvest diseases on oranges.     

Molecular Approaches to Assist the Screening and Monitoring of Postharvest Biocontrol Yeasts

Nineteen yeast isolates obtained from the surface of several fruits and vegetables grown in Southern Italy and Israel were compared by molecular analysis using arbitrarily primed polymerase chain reaction (AP-PCR) and random amplified polymorphic DNA technique (RAPD-PCR). Genetic analysis made it possible to distinguish between closely-related genitically different strains which had the same morphological characteristic, and to discard isolates which were genetically identical. Following PCR characterisation, 6 isolates were selected and tested for their biocontrol activity against major postharvest pathogens (Penicillium digitatum on grapefruit, Botrytis cinerea, Rhizopus stolonifer and Aspergillus niger on table grape and B. cinerea and R. stolonifer on cherry tomato). All the isolates showed a good biocontrol efficacy on both wounded and non-wounded fruits. Furthermore, the preharvest application of the most effective antagonist (LS15) on table grape resulted in a significant reduction in grey mold ranging from 28.3% to 38.2% compared to the untreated control. The RAPD-PCR technique was also useful for identifying and monitoring the survival of the antagonist after field application.     

柑桔贮藏病害发生动态和化学防腐保鲜技术的研究

作者1983～1995年间研究了浙江自然状态下柑桔贮藏病害的发生动态。结果显示,以采后侵染为主的病害发病率占59.79±22.40％;采前采后均可侵染,但以采前侵染为主的病害发病率占40.21±22.40％。后一类病害为害加剧是浙江柑桔贮藏病害发生的新动态,从国内外16种药剂中筛选出抑霉唑(Imazalil)、扑霉灵(Prochloraz)、百可得(Bellkute)等新型柑桔防腐保鲜剂,大量用于生产,解决了柑桔青、绿霉病菌对苯并咪唑类药剂的抗药性问题,经济和社会效益显著。研究了抑霉唑对柑桔青、绿霉病菌的敏感基线,在国内外首次报道大幅度降低使用浓度(仅原来的1/2～1/4)仍有良好效果,并率先在浙江桔区大量使用。     

Characterisation of the fungal population in citrus packing houses

The aim of this work was the characterisation of the environmental and superficial mycoflora of equipment and facilities of two citrus packing houses in Sao Paulo State, Brazil, in 2004 and 2005. One of the packing houses packed fruit for the export market (municipality of Matao), the other for the domestic market (municipality of Engenheiro Coelho). The study also identified the presence of isolates of Penicillium spp. resistant to thiabendazole and imazalil fungicides in packing houses. The environmental mycoflora was sampled according to the gravimetric method, using Petri dishes containing potato dextrose agar medium opened for 2 min. The superficial mycoflora on equipment and facilities was sampled with Rodac plates. The mycoflora in the environment and on surfaces of the packing houses in Matao were 12.3 and 52.3 cfu/plate, respectively, while these populations for the Engenheiro Coelho packing house were 46.3 and 68.2 cfu/plate, respectively. Cladosporium and Penicillium were the most prevalent genera of fungi. The contamination levels of clean zones in the packing houses (washing of fruits, packing table, boxes and containers) was not substantially lower than the contamination in dirty zones (reception of fruits and first selection). The percentage of P. digitatum isolates in Matao that was resistant to thiabendazole and imazalil was 25.9 and 1.5 in the environment and 30.1 and 16.0 on packing house surfaces, respectively. In Engenheiro Coelho, percentage of resistance to these fungicides was 51.9 and 0.1 in the environment and 39.2 and 0.9 on packing house surfaces, respectively.     

Production of hydrogen peroxide and expression of ROS‐generating genes in peach flower petals in response to host and non‐host fungal pathogens

Reactive oxygen species (ROS) play dual roles in plant–microbe interactions in that they can either stimulate host resistance or enhance pathogen virulence. Innate resistance in peach (Prunus persica) to the brown rot fungal pathogen Monilinia fructicola is very limited, and knowledge of the mechanism of virulence is rudimentary. In this study, production of hydrogen peroxide, a major component of ROS, was determined in peach flower petals in response to M. fructicola (a host pathogen) and Penicillium digitatum (a non‐host pathogen). Monilinia fructicola was able to infect flower petals while P. digitatum was not. During the host‐specific interaction, M. fructicola induced hydrogen peroxide accumulation in flower petals. Application of exogenous antioxidants significantly reduced hydrogen peroxide accumulation as well as the incidence of brown rot disease. Application of M. fructicola spores to the surface of intact flower petals induced gene expression and increased enzyme activity of NADPH oxidase and cell wall peroxidase in host tissues, resulting in the production of hydrogen peroxide. Petals inoculated with M. fructicola exhibited high levels of protein carbonylation and lipid peroxidation. No significant response in gene expression, enzyme activity or hydrogen peroxide levels was observed in peach flower petals treated with P. digitatum. These results suggest that M. fructicola, as with other necrotrophic fungi, uses the strong oxidative response as part of a virulence mechanism.     

寡雄腐霉GAQ1对辣椒疫病的防效及对辣椒的促生作用

为明确自主分离的生防菌株寡雄腐霉Pythium oligandrum GAQ1对辣椒疫病的生防效果及其防御机制,通过平板拮抗和盆栽防效试验测定寡雄腐霉菌株GAQ1对辣椒疫霉Phytophthora capsici菌丝的拮抗作用、对辣椒疫病的防效和对辣椒的促生效果,同时应用实时荧光定量PCR技术检测菌株GAQ1处理后辣椒抗性基因表达的变化。结果表明,寡雄腐霉菌株GAQ1的菌丝可以缠绕并吸附寄生在辣椒疫霉菌丝表面或穿入菌丝体内,使辣椒疫霉菌丝细胞死亡;菌株GAQ1发酵液处理辣椒离体叶片再接种辣椒疫霉后产生的病斑直径较对照组显著减少,离体防效为30.79%;接种菌株GAQ1菌丝球后,辣椒疫病的病情指数较对照组显著降低,盆栽防效达69.16%;经菌株GAQ1处理辣椒后可诱导相关抗性基因PR1、WRKY40、WRKY53、ACCO和GST的相对表达量出现不同程度的升高,说明菌株GAQ1可诱导辣椒植株产生不同程度的防御系统应答。菌株GAQ1对辣椒具有良好的促生效果,处理后第5周其株高、株重及根重分别较对照组提高10.11%、33.23%和24.72%,其叶片中叶绿素a、叶绿素b及类胡萝卜素的含量分...     

解淀粉芽胞杆菌EA19菌株对小麦赤霉病的防治效果

为明确解淀粉芽胞杆菌Bacillus amylolyticus EA19菌株对小麦赤霉病的防治效果,室内测定EA19菌株及其发酵液对小麦赤霉病菌Fusarium asiaticum菌落生长、孢子萌发、子囊壳形成的影响,以及在室内离体麦穗上的防治效果,并进行了田间小区防治试验。室内试验测定结果表明,EA19菌株发酵液对小麦赤霉病菌菌落生长和分生孢子的萌发具有显著的抑制作用,在室内离体麦穗上的防治效果可达80.0%,对子囊壳形成无显著影响。田间小区防治试验结果表明,在添加紫外保护剂抗坏血酸的条件下,EA19菌株发酵液和菌体悬浮液对赤霉病均有显著的防治效果,其中EA19菌株发酵液的防治效果可达81.2%。EA19菌株喷雾粉可湿性粉剂对赤霉病的防治效果可达67.4%,能使小麦籽粒中脱氧镰刀菌烯醇毒素含量显著降低36.8%,与50%多菌灵可湿性粉剂的效果无显著差异。表明EA19菌株在生产上防治赤霉病具有较大的开发应用潜力。     

Modes of Action of Pantoea agglomerans CPA-2, an Antagonist of Postharvest Pathogens on Fruits

Pantoea agglomerans CPA-2 is an effective antagonist against the postharvest pathogens Penicillium digitatum and Penicillium italicum on citrus fruits but its mode of action is unknown. Possible mechanisms studied in this work were antibiosis, induced resistance, competition and production of chitinolytic enzymes. P. agglomerans CPA-2 was unable to produce antibiotics or chitinolytic enzymes under the conditions tested. Induction of resistance by P. agglomerans CPA-2 was studied in oranges by measuring phenylalanine ammonia lyase and peroxidase enzyme activity in the orange peel at different time points after inoculation with the antagonist and/or the pathogen. No significant augmentation of enzyme activity after inoculation of oranges with P. agglomerans CPA-2 in the presence or absence of the pathogen was observed. P. agglomerans was effective only when it is in close contact with the pathogens. Competition for nutrients was studied using tissue culture plates with cylinder inserts, which allowed competition for nutrients to be studied without competition for space since physical contact between pathogen and antagonist was avoided. The presence of P. agglomerans in the tissue culture wells clearly decreased the germination of Penicillium conidia present in the cylinder when diluted orange peel extract or diluted potato dextrose broth was the nutrient source. Germination of Penicillium conidia, however, was almost completely inhibited when pathogen and antagonist were in physical contact. These results indicate that competition for nutrients is one of the modes of action of P. agglomerans CPA-2, but that physical contact between pathogen and antagonist is important for effective control.     

Control of postharvest green mould of citrus by haliscosamine isolated from Haliclona viscosa sponge

The aim of this study was to evaluate the antifungal activity of haliscosamine isolated from Haliclona viscosa sea sponge from the Moroccan Atlantic coast against Penicillium digitatum, the causal agent of citrus green mould. The in vitro antifungal activity of haliscosamine, determined using the agar disc‐diffusion method, showed that haliscosamine and DESOGERME SP VEGETAUX^® (reference product) have fungicidal activity with inhibition diameters of 19 and 18 mm, respectively. Haliscosamine and a positive control (DESOGERME SP VEGETAUX^®) were tested in vivo in comparison with a negative control on orange fruits in normal storage conditions. This test showed that after four weeks of storage, the rate of decay of citrus fruits treated with haliscosamine or DESOGERME does not exceed 14%, whereas in the negative control the decay rate is 77%. The results of in vitro and in vivo tests indicate that haliscosamine and DESOGERME SP VEGETAUX^® have an important antifungal potential against P. digitatum and that haliscosamine (a natural product) may represent an alternative to the use of chemical fungicides.     

Optimised expression and spectral analysis of the target enzyme CYP51 from Penicillium digitatum with possible new DMI fungicides

BACKGROUND: Sterol 14α‐demethylase (CYP51), a key target of azole (DMI) fungicides, can be expressed in both prokaryotes and eukaryotes. Green mould of citrus, caused by Penicillium digitatum (Pers.) Sacc., is a serious post‐harvest disease. To develop specific and more effective fungicides against this disease, the characteristics of the interaction between sterol 14α‐demethylase from P. digitatum (PdCYP51) and possible new fungicides were analysed. The cyp51 gene of P. digitatum was cloned and expressed under different conditions in Escherichia coli (Mig.) Cast. & Chalm., and the binding spectra of PdCYP51 were explored by the addition of two commercial azoles and four new nitrogen compounds. RESULTS: The yield of soluble protein (PdCYP51) was largest when expressed in Rosetta (DE3) induced by 0.5 mM IPTG for 8 h at 30 °C. Compound B (7‐methoxy‐2H‐benzo[b][1,4]thiazine‐3‐amine) showed the strongest binding activity of the four new nitrogen compounds, with a K_d value of 0.268 µM . The K_d values of the six compounds were significantly correlated with their EC₅₀ values. CONCLUSION: The spectral analysis and bioassay results could be used to screen the new chemical entities effectively. Compound B, selected by virtual screening from a commercial chemical library, is a candidate for a new DMI fungicide. These results provide a theoretical basis and new ideas for efficient design and development of new antifungal agents. Copyright © 2010 Society of Chemical Industry     

Control of<Emphasis Type="Italic">Penicillium digitatum</Emphasis> on citrus fruit using two plant extracts and study of their mode of action

Extracts from two plants from Ethiopia,Withania somnifera andAcacia seyal, were evaluated for their potential as natural biopesticides and to study their mode of action. Methanolic extracts of these plants were testedin vivo on citrus fruit for their efficacy to controlPenicillium digitatum when applied on wounded or unwounded fruit surfaces. Relative to the control, 70% and 75% of wound — inoculated fruit did not develop decay symptoms for up to 21 days of storage at 25°C and >85% r.h. An increase in cell wall-bound phenolics was evident in wounded fruit treated with plant extracts and inoculated with a spore suspension ofP. digitatum. Scanning electron microscopy revealed deposition of crystalline plant material sticking to the pathogen and around the wound site. The application of the plant extracts increased the epiphytic background total microbial population but decreased diversity. http://www.phytoparasitica.org posting May 7, 2007.     

Induction of Chitinase and β-1,3-Endoglucanase Proteins by UV Irradiation and Wounding in Grapefruit Peel Tissue

UV irradiation enhanced the resistance of grapefruit against the development of green mold décay caused byPenicillium digitatum, the main postharvest pathogen of citrus fruit, and significantly inhibited the fungus’ growth at the fruit wound sites. Immunoblotting analysis using specific citrus chitinase and β-1,3-endoglucanase antibodies, showed that UV irradiation, wounding of the fruit, or a combination of these two treatments, induced the accumulation of a 25 kD chitinase protein in the fruit’s peel tissue. On the other hand, UV irradiation or wounding of the fruit alone was unable to induce the accumulation of 39 and 43 kD β-1,3-endoglucanase proteins, but the combination of the two treatments increased these protein levels. It is suggested that both chitinase and β-1,3-endoglucanase may play a role in the UV-induced resistance of grapefruit againstP. digitatum. Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. 403/99. http://www.phytoparasitica.org posting June 3, 1999.     

泽兰实蝇寄生对紫茎泽兰生长和防御能力的影响

为明确泽兰实蝇Procecidochares utilis寄生对紫茎泽兰Ageratina adenophora生长及防御能力的影响,在温室内设置泽兰实蝇寄生与未寄生紫茎泽兰2组盆栽处理,测定泽兰实蝇寄生对紫茎泽兰生长特性、生物量、叶片营养物质、次生代谢物质和保护酶的影响。结果表明,泽兰实蝇寄生显著降低了紫茎泽兰的株高、茎粗、叶片厚度和叶片数,分别显著降低了24.80%、16.86%、42.31%和27.82%;同时,地上生物量、根部生物量和总生物量分别显著降低了58.89%、54.29%和57.60%。泽兰实蝇寄生还导致紫茎泽兰的营养成分发生显著变化,其中可溶性蛋白、可溶性糖、可溶性淀粉和叶绿素含量分别显著降低了41.18%、11.75%、34.57%和29.96%。泽兰实蝇寄生改变了紫茎泽兰的保护酶活性,其过氧化氢酶和超氧化物歧化酶活性分别比未寄生植株显著升高了59.36%和108.06%,过氧化物酶活性降低了25.69%;同时使紫茎泽兰单宁酸含量显著降低了21.67%,总酚和类黄酮的含量分别显著上升了1.21倍和2.09倍。泽兰实蝇寄生的紫茎泽兰总生物量与次生代谢物质总酚和类黄酮的含量呈显著负相关,相关系数r分别为-0.06和-0.43,对照组则不存在相关性。表明泽兰实蝇寄生可能促使紫茎泽兰改变自身的生长与防御能量分配,减少了营养和生长的投入,将更多的资源投向繁殖和防御。     

撕裂蜡孔菌对烟草黑胫病的防治效果及对烤烟生长的影响

为探讨撕裂蜡孔菌Ceriporia lacerata菌株HG2011对烟草黑胫病的防治效果和对烤烟的促生作用,利用该菌株制备发酵液和固体菌剂,通过拮抗和盆栽试验研究其对烟草黑胫病病原菌烟草疫霉Phytophthora nicotianae生长的抑制作用、对黑胫病的防治效果及对烟苗生长和养分吸收的影响。结果表明:撕裂蜡孔菌菌株HG2011能显著抑制烟草疫霉生长;在带毒平板试验中,含20%发酵液的培养基对烟草疫霉菌丝生长的抑制率达54.50%;在平板对峙试验中,菌株HG2011能使烟草疫霉菌丝扭曲、畸形和空泡化,最终完全侵入并覆盖烟草疫霉菌落。在盆栽试验中,施用撕裂蜡孔菌固体菌剂可降低发病烟苗叶片中的丙二醛含量,并能维持正常的细胞膜透性,使烟苗黑胫病发病率比单独接种烟草疫霉处理显著降低了15.55%～37.77%,且预防效果71.16%优于治疗效果17.98%。在化肥配施高量菌剂处理中烟苗的株高、最大叶面积和生物量均最高,比单施化肥处理显著增加9.08%、36.28%和11.84%;烟苗氮、磷、钾的吸收量比单施化肥处理显著增加11.29%、38.73%和12.79%。表明撕裂蜡孔菌菌株HG2011能有效防治烟草黑胫病,刺激烟苗生长、促进养分吸收,表现出良好的防病促生效应。     

壳寡糖对干旱胁迫下信前胡生长及叶片生理特性的影响

以信前胡盆栽幼苗作为研究材料,通过测定信前胡的株高、叶面积、生物量、含水量、根冠比和叶片抗旱相关的生理指标,研究了5、10、15 mg·L^-1和2 mg·L^-1的壳寡糖(Chitooligosaccharide,COS)溶液处理对干旱胁迫下信前胡生长和叶片生理特性的影响。结果表明,不同浓度壳寡糖对干旱胁迫下信前胡生长均有不同程度的促进作用,且随壳寡糖浓度的增加,各生长指标均表现出先增后降的趋势,均有一定的最适浓度。与单纯干旱处理相比,5、10、15 mg·L^-1和20 mg·L^-1壳寡糖处理信前胡的单株叶片数分别提高了32.5%、50.8%、29.3%和6.3%,均达到了显著水平（P<0.05）;10 mg·L^-1和15 mg·L^-1壳寡糖处理信前胡的单株平均叶面积分别提高了27.3%和20.8%,均达到了显著水平（P<0.05）;5、10、15 mg·L^-1壳寡糖处理信前胡的株高分别显著提高了17.4%、31.7%和19.6%。10 mg·L^-1壳寡糖处理信前胡叶片的脯氨酸含量较单纯干旱处理显著提高了11.8%,10 mg·L^-1壳寡糖处理信前胡叶片的H₂O₂含量和MDA含量均较单纯干旱处理显著降低（降幅分别为46.9%和14.4%）。因此,10 mg·L^-1壳寡糖处理可显著缓解信前胡苗期的干旱胁迫（P<0.05）。